RNA Methodologies Laboratory Guide for Isolation and Characterization

ISBN-13: 9780128046784 / Angielski / Spiral bound / 2017 / 876 str.

Farrell, Robert E., Jr. (Pennsylvania State University, York, PA, USA)

Opis

RNA Methodologies, Fifth Edition continues its tradition of excellence in providing the most up-to-date ribonucleic acid lab techniques for seasoned scientists and graduate students alike. This edition features new material on the exploding field of microRNA as well as the methods for the profiling of gene expression, both which have changed considerably in recent years. As a leader in the field, Dr. Farrell provides a wealth of knowledge on the topic of RNA while also giving readers helpful hints from his own personal experience in this subject area. Beginning with the most contemporary, RNA Methodologies, Fifth Edition, presents the essential techniques to use when working with RNA for the experienced practitioner while at the same time providing images and examples to aid the beginner in fully understanding this important branch of molecular biology. The next generation of scientists can look to this work as a guide for ensuring high productivity and highly representative data, as well as best practices in troubleshooting laboratory problems when they arise.

• Features new material in miRNA, MIQE guidelines, biomarkers, RNA sequencing, digital PCR and more
• Includes expanded coverage on quantitative PCR techniques, RNAi, bioinformatics, the role of locked nucleic acids, aptamer biology, PCR arrays, and other modern technologies
• Presents comprehensive, cutting-edge information covering all aspects of working with RNA
• Builds from basic information on RNA techniques to in-depth protocols to guidance on how to modify and adjust each step of a particular application
• Presents multiple avenues for addressing the same experimental goals

Spis treści

1. RNA and the Cellular Biochemistry Revisited2. Creating a Ribonuclease-free Environment3. RNA Isolation Strategies4. The Truth about Tissues5. Going Green: RNA and the Molecular Biology of Plants6. Quality Control for RNA Preparations7. cDNA: A Permanent Biochemical Record of the Cell8. RT-PCR: A Science and an Art Form9. Quantitative PCR Techniques10. miRNA11. RNA Interference - Take a RISC: Role the Dicer12. Stringency: Conditions that Influence Nucleic Acid Structure13. Electrophoresis of RNA14. Photodocumentation and Image Analysis15. Northern Analysis16. Nucleic Acid Probe Technology17. Isolation of Polyadenylated RNA18. Quantification of Specific mRNAs by Nuclease Protection19. Analysis of Nuclear RNA20. Non-Array Methods for Global Analysis of Gene Expression21. Genomes, Transcriptomes, Proteomes, and Bioinformatics22. RNA Biomarker Development23. High-Throughput Analysis of Gene Expression24: Functional Genomics and Transcript Profiling25. A Few RNA Success Stories

AppendixA. Maintaining Complete and Accurate RecordsB. Converting Mass to MolesC. Disposal of Ethidium Bromide and SYBR Green SolutionsD. DNase I Removal of DNA from an RNA SampleE. RNase Incubation to Remove RNA from a DNA SampleF. Useful Stock Solutions for the Molecular BiologistG. Silanizing Centrifuge Tubes and GlasswareH. Dot Blot AnalysisI. Electrophoresis: Principles, Parameters, and SafetyJ. Polyacrylamide Gel ElectrophoresisK. Centrifugation as a Mainstream Tool for the Molecular BiologistL. Trypsinization Protocol for Anchorage-Dependent CellsM. Isolation of High-Molecular-Weight DNA by Salting-OutN. Phenol PreparationO. Deionization of Formamide, Formaldehyde, and GlyoxalP. Selected Suppliers of Equipment, Reagents, and ServicesQ. Useful SI UnitsR. Common AbbreviationsS. Trademark Citations